Laboratory of Cellular and Molecular Biology, Department of Clinical Pathology, National Institute of Gastroenterology, "S. De Bellis" Research Hospital, Via Turi 27, 70013, Castellana Grotte, BA, Italy.
Program for Targeted Experimental Therapeutics, Izmir Biomedicine and Genome Center, Dokuz Eylul University, Izmir, Turkey.
Cell Oncol (Dordr). 2018 Jun;41(3):283-296. doi: 10.1007/s13402-018-0370-z. Epub 2018 Feb 22.
Emerging evidence indicates that combining Sorafenib with vitamin K1 (VK1) may result in a synergistic inhibition of hepatocellular carcinoma (HCC) cell migration and proliferation. Despite this synergy, its benefits may be limited due to drug resistance resulting from cross-talk with the tumor microenvironment. Insulin-like growth factor-1 (IGF1) signaling acts as an important modulator of HCC cell growth, motility and drug resistance. Therefore, we aimed to explore the effects of Sorafenib in combination with VK1 and/or IGF1-R antagonists on HCC cells.
Scratch wound migration assays were performed to assess the motility of HCC-derived PLC/PRF/5, HLF and Hep3B cells. The synergistic, additive or antagonistic effects of Sorafenib, VK1 and IGF1-R antagonists on HCC cell motility were assessed using CompuSyn software. The effects mediated by these various compounds on HCC cytoskeleton organization were evaluated using DyLight 554 Phalloidin staining. Proliferation and migration-associated signaling pathways were analyzed in PLC/PRF/5 cells using Erk1/2 and Akt activation kits and Western blotting (Mek, JNK, Akt, Paxillin and p38), respectively.
The effects of the IGF1-R antagonists GSK1838705A and OSI-906 on HCC cell migration inhibition after Sorafenib and/or VK1 administration, individually or in combination, were evaluated. We found a synergistic effect in PLC/PRF/5, HLF and Hep3B cells for combinations of fixed doses of GSK1838705A or OSI-906 together with different doses of Sorafenib and/or VK1. The levels of synergy were found to be stronger at higher Sorafenib and/or VK1 concentrations and lower or absent at lower concentrations, with some variation among the different cell lines tested. In addition, we found that in PLC/PRF/5 and HLF cells IGF1-R blockage strongly enhanced the reduction and redistribution of F-actin induced by Sorafenib and/or VK1 through alterations in the phosphorylation levels of some of the principal proteins involved in the MAPK signaling cascade, which is essential for cell migration.
Our results indicate that modulation of the efficacy of Sorafenib through combinations with VK1 and/or IGF1-R antagonists results in synergistic inhibition of HCC cell migration.
新出现的证据表明，索拉非尼与维生素 K1（VK1）联合使用可能会导致肝细胞癌（HCC）细胞迁移和增殖的协同抑制。尽管存在这种协同作用，但由于与肿瘤微环境的相互作用导致的耐药性，其益处可能会受到限制。胰岛素样生长因子-1（IGF1）信号作为 HCC 细胞生长、迁移和耐药性的重要调节剂。因此，我们旨在探讨索拉非尼与 VK1 和/或 IGF1-R 拮抗剂联合应用对 HCC 细胞的影响。
划痕迁移实验用于评估源自 PLC/PRF/5、HLF 和 Hep3B 细胞的 HCC 细胞的迁移能力。使用 CompuSyn 软件评估索拉非尼、VK1 和 IGF1-R 拮抗剂对 HCC 细胞迁移的协同、相加或拮抗作用。使用 DyLight 554 鬼笔环肽染色评估这些化合物对 HCC 细胞骨架组织的影响。使用 Erk1/2 和 Akt 激活试剂盒和 Western blot（Mek、JNK、Akt、Paxillin 和 p38）分别分析 PLC/PRF/5 细胞中增殖和迁移相关信号通路。
评估了 IGF1-R 拮抗剂 GSK1838705A 和 OSI-906 在单独或联合使用索拉非尼和/或 VK1 后对 HCC 细胞迁移抑制的影响。我们发现，在 PLC/PRF/5、HLF 和 Hep3B 细胞中，GSK1838705A 或 OSI-906 的固定剂量与不同剂量的索拉非尼和/或 VK1 联合使用时具有协同作用。在较高的索拉非尼和/或 VK1 浓度下，协同作用的水平更强，而在较低的浓度下则较低或不存在，不同测试的细胞系之间存在一些差异。此外，我们发现，在 PLC/PRF/5 和 HLF 细胞中，IGF1-R 阻断通过改变 MAPK 信号级联中一些主要蛋白的磷酸化水平，强烈增强了索拉非尼和/或 VK1 诱导的 F-肌动蛋白的减少和重分布，这对于细胞迁移是必需的。
我们的结果表明，通过与 VK1 和/或 IGF1-R 拮抗剂联合使用来调节索拉非尼的疗效，可导致 HCC 细胞迁移的协同抑制。
